• Bulletin of the Korean Chemical Society
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• v.11 no.4
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• pp.297-302
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• 1990

Retention of mono-substituted phenols in reversed-phase liquid chromatography has been studied based on the linear solvation energy relationships using the solvatochromic mobile phase parameters, ${\pi}_m^{\ast}, {\alpha}_m$ and ${\beta}_m$. It has been observed that retention behavior of phenols in RPLC were well represented by regression equations vs. solvatochromic mobile phase parameters even though the equations may be incomplete due to lack of an explicit cavity term. Dependence of retention of monosubstituted phenols on the mobile phase properties were varied depending on the type of the organic cosolvent in the mobile phase, e.g., ${\beta}_m$ and {\alpha}_m$ in methanol-water system, but ${\pi}_m^{\ast} and ${\beta}_m$ in THF-water system. It has been suggested that retention of phenols in methanol-water system is controlled by the solvophobicity of the mobile phase.

• Analytical Science and Technology
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• v.7 no.3
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• pp.379-386
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• 1994

The elution behaviors in reversed-phase liquid chromatography were investigated thermodynamically for 2-hydroxy-arylazopyrazolone chelates with Ni(II), Cu(II), Co(III), Cr(III) on Novapak $C_{18}$ column. There was a good linear dependence of the capacity factor(k') on the variations of column temperature in van't Hoff plot. From this result, it was confirmed that the retention mechanism of these chelates in the reversed phase liquid chromatography system was invariant under the condition of various temperatures. For the most cases of the chelates studied, the dependence of capacity factor(1n k') on enthalpy$(-{\Delta}H)^{\circ}$, calculated by van't Hoff plot showed a good linearity(r=0.980~0.999) except [Pm(2-OH_(5-Cl)PaPz](r=0.787) and also the compensation temperatures(${\beta}$) showed constant values. The range of compensation temperature values calculated from the slope of $-{\Delta}H^{\circ}$ vs 1n k' plots was 374.3~806.9K. It was suggested that the retention of metal-2-hydroxy-arylazopyrazolone chelates in the reversed phase liquid chromatography system was largely affected by the hydrophobic effect.

• Analytical Science and Technology
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• v.13 no.3
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• pp.399-402
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• 2000

To determine the concentrations of microcystins present in lake water or in tap water using high performance liquid chromatography, it is necessary to concentrate a large volume of water samples (about 20 L) into very small volume (0.1-0.3 mL). Concentration can be conveniently done when disc type solid phase extraction (SPE) apparatus is used. Using this apparatus we have investigated the recovery rates of three kinds of microcystins, RR, YR, LR. The recovery rates were relatively low and the reproducibilities were not good either. It is expected, however, that the appropriate selection of the disc conditioning and eluting solvents and reproducible reconcentration process after SPE will improve both the recovery rates and the reproducibilities.

• FOCUS: LIFE SCIENCE
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• no.1
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• pp.6.1-6.9
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• 2024

The document is a white paper on Hydrophilic Interaction Liquid Chromatography (HILIC) analysis method development. HILIC is a type of chromatography that uses an organic/aqueous mobile phase and a polar stationary phase. In HILIC, water is a strong solvent, and unlike in Reversed Phase Liquid Chromatography (RPLC), increasing the proportion of water in the mobile phase reduces the retention time of the analyte. The paper discusses when to consider HILIC analysis methods, the advantages of HILIC, and the challenges often encountered due to the lack of understanding of HILIC mechanisms compared to RPLC. It also provides a systematic flowchart for intelligent solutions for HILIC analysis method development, which includes a three-step approach for chromatography analysis method development. The first step involves gathering as much information as possible about the analyte (e.g., pKa, log P, log D). The second step involves analyzing the sample under different pH conditions using three HILIC columns in either isocratic or gradient mode to identify the suitable column/pH combination for the analyte. The third step involves optimizing the separation by investigating other parameters such as temperature and ionic strength, and assessing the robustness of the method. The paper emphasizes that the selection of the appropriate stationary/mobile phase combination, based on the differences between the HILIC stationary phases and the mobile phase pH, can provide high selectivity in the analysis. This step-by-step approach can help users develop an efficient analysis method.

• Food Science of Animal Resources
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• v.40 no.1
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• pp.87-95
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• 2020

In this study, we separated and purified lipase inhibitory peptide from fermented milk by Lactobacillus plantarum Q180 with the aim of developing a new functional anti-lipase activity yogurt product. L. plantarum 180 was inoculated into 10% reconstituted skimmed milk and incubated at 37℃ until the pH of the culture reached pH 4.4. The lipase activity was measured using porcine pancreatic lipase. The lipase inhibitory peptides were gradually isolated by ultrafiltration, reversed phase column chromatography (RPC), reversed phase high-performance liquid chromatography (RP-HPLC), and gel permeation high-performance liquid chromatography (GP-HPLC) from the fermented milk by L. plantarum Q180. An ODS-AQ column was used for the RPC, a Vydac C18 column for the RP-HPLC, and a Superdex Peptide HR column for the GP-HPLC. The peptide was composed of Asp, Thr, Ile, Ser, Ala, and Gln, and the anti-lipase activity (IC₅₀) was 2,817 ㎍/mL.

• Bulletin of the Korean Chemical Society
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• v.11 no.6
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• pp.568-570
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• 1990

• Bulletin of the Korean Chemical Society
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• v.9 no.6
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• pp.341-345
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• 1988

Separation of optical isomers of dansyl amino acids by a reversed phase liquid chromatography has been accomplished by adding a copper (II) chelate of N-benzyl-L-proline to the mobile phase. The pH, the eluent composition and the concentration of copper (II) chelate all affect the optical separations. The elution orders between D and L DNS-amino acids were consistant except dansyl phenylalanine showing that D forms of DNS-amino acids elute earlier than L forms. These behaviors are different from the results obtained by the use of copper (II) proline. The retention mechanism for the optical separation of the dansyl amino acids can be explained by the equilibrium of liqand exchange and by hydrophobic interaction.

• Analytical Science and Technology
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• v.5 no.1
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• pp.63-71
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• 1992

Liquid chromatographic behavior of Pd(II), Ni(II) and Co(III) in N-Alkylisonitrosoacetylacetone imine(HIAA-NR) chelates was investigated by reversed phase high perfomance liquid chromatography. The optimum conditions for the separation of IAA-NR-metal chelates were examined respect to the flow rate and mobile phase strength. The metal-N-Alkylisonitrosoacetylacetone imine chelates in solution were successfully separated on Novapak $C_{18}$ column using acetonitrile/water mixture as mobile phase. The elution order of chelates is methyl>ethyl>propyl>butyl as N-alkyl group for ligand is varied. It was found that all IAA-NR-metal chelates were eluted in an acceptable range of capacity factor value($0{\leq}log\;k^{\prime}{\leq}1$). The dependence of log k' on the volume fraction of water in the binary mobile phase was examined. Also, the dependence of k' on the liquid-liquid extraction distribution ratio(Dc) in acetonitrile-water-alkane extraction system was investigated for IAA-NR-metal chelate. Both kinds of dependence are linear, which suggests that the retention of the electroneutral metal chelates on Novapak $C_{18}$ column is largely due to the hydrophobic effect.

• Bulletin of the Korean Chemical Society
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• v.11 no.5
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• pp.411-415
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• 1990

The dabsylation of amino acids has been applied to resolve their optical isomers with the use of chiral mobile phase in high performance liquid chromatography. The dabsyl amino acids were successfully separated on reversed phase column($C_{18}$) by adding a chiral L-benzylproline-Cu(II) chelate to the mobile phase. The separation selectivity of the dabsyl amino acid enantiomers was not less than that of dansyl amino acids. The retention order of the dabsyl amino acid enantiomers was as those of the dansyl amino acid enantiomers except dabsyl threonine. The optical selectivity of the dabsyl amino acids increase with pH of the mobile phase and concentration of the chelate, but slightly decreases with concentration of buffer and organic solvent composition. However serine, methionine, valine, and leucine showed a slight decrease in the optical selectivity with increase in pH. The retention times of the dabsyl amino acids decreases with increasing pH and acetonitrile concentration but increases with the concentration of the chiral chelate added. The mechanism of the optical resolution is based on a stereospecific interaction including a intramolecular hydrophobic effect and SN-2 reactivity of the ligand exchange chromatography.It is advantageous to detect absorption at 436 nm, which is less interferent them the other detection systems. The derivatized dabsyl amino acids are stable for a month.

• Bulletin of the Korean Chemical Society
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• v.28 no.4
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• pp.601-606
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• 2007

The chromatographic behaviors of nucleotides (inosine 5'-monophosphate, uridine 5'-monophosphate, guanosine 5'-monophosphate, and thymine monophosphate disodium salts) on a C18 column were studied with different types of ionic liquids (ILs) as additives for the mobile phase in reversed-phase liquid chromatography (RPLC). Three ILs, 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIm][BF4]), 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIm][BF4]), and 1-ethyl-3-methylimidazolium methylsulfate ([EMIm][MS]), were used. Eluents were composed of water and methanol (90/10%, vol) with the addition of 0.5-13.0 mM of ILs. The effects of the concentration of ILs on retention and separation were investigated and discussed. The results showed that the addition of ILs affects the retention and resolution of the tested compounds. Use of 13.0 mM of [BMIm][BF4] as the eluent modifier resulted in a baseline separation of nucleotides without requiring gradient elution. This study demonstrates that ILs can be potentially applied as a mobile phase modifier in RPLC.